LOCUS       sequence-207881-        6958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NFS1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6958)
  AUTHORS   Alvarez SW, Sviderskiy VO, Terzi EM, Papagiannakopoulos T, Moreira 
            AL, Adams S, Sabatini DM, Birsoy K, Possemato R
  TITLE     NFS1 undergoes positive selection in lung tumours and protects cells
            from ferroptosis.
  JOURNAL   Nature. 2017 Nov 22. doi: 10.1038/nature24637.
  PUBMED    29168506
REFERENCE   2  (bases 1 to 6958)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2017 Nov 22. doi: 10.1038/nature24637."
COMMENT     Sequence Label: pBabe hygro NFS1
FEATURES             Location/Qualifiers
     source          1..6958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             98..567
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    632..831
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1032..1448
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1378..1400
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1416..1432
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2881..2901)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2886..3215
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3066..3201
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3289..4314
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3289..4314
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     4831..4850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5112..5581
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     gap             6000..6010
                     /estimated_length=11
     CDS             complement(join(6060..6851,6852..6920))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6683..6702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6921..6958,1..67))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
       61 ttccgcgcac atttccccga aaagtgccac ctgcagcctg aatatgggcc aaacaggata
      121 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
      181 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
      241 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
      301 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
      361 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
      421 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
      481 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      541 tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg
      601 cccagggacc accgacccac caccgggagg caagctggcc agcaacttat ctgtgtctgt
      661 ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag ttagctaact
      721 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
      781 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
      841 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
      901 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
      961 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     1021 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1081 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1141 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1201 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1261 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1321 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1381 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1441 taggcgccgg ccggatccac catgctgctc cgagccgctt ggaggcgggc ggcagtggcg
     1501 gtgacagcgg ctccagggcc gaagcccgcg gcgcccactc gggggctgcg cctgcgcgtt
     1561 ggagaccgtg ctcctcagtc tgcggttccc gcagatacag ccgctgcccc ggaggtgggg
     1621 ccagtgctgc gacctctcta tatggatgtg caagctacaa ctcctctgga cccccgggtg
     1681 cttgatgcca tgctccctta cctaattaac tactatggga acccacactc ccggacacat
     1741 gcttatggct gggagagtga ggcagccatg gaacgtgctc gtcagcaagt agcatctctg
     1801 attggagctg atcctcgtga gatcattttt actagtggtg ctactgaatc caacaacata
     1861 gcaattaagg gggtggcccg attctacagg tcacggaaaa agcacttgat caccacccag
     1921 acagaacaca aatgtgtctt ggactcctgc cgttcactgg aagctgaggg ctttcaggtc
     1981 acctacctcc cagtgcagaa gagtgggatc attgacctaa aggaactaga ggctgctatc
     2041 cagccagata ctagcctggt gtcagtcatg actgtgaaca atgagattgg agtgaagcag
     2101 cctattgcag aaatagggcg gatttgttca agtcgtaaag tttacttcca tactgatgca
     2161 gcccaggctg ttggaaaaat cccacttgat gtcaatgaca tgaaaattga tctcatgagc
     2221 attagtggtc acaaaatcta cggtcccaaa ggggttggtg ccatctacat ccgtcgccgg
     2281 ccccgtgtgc gtgtggaggc cctgcagagt ggaggggggc aggagcgggg tatgcggtct
     2341 gggacagtgc ccacaccctt agtggtgggg ctgggggctg cgtgtgaggt ggcacagcaa
     2401 gagatggagt atgaccacaa gcgaatctca aagttgtcag agcggctgat acagaatata
     2461 atgaagagcc ttccagatgt ggtgatgaat ggggacccta agcaccatta tcccggctgt
     2521 atcaacctct cctttgcata tgtggaaggg gaaagtctgc tgatggcact gaaggacgtt
     2581 gccttatcct cagggagtgc ctgcacctct gcatccctgg agccctctta tgtgcttaga
     2641 gcaattggca ctgatgagga tttagcgcac tcttctatca ggtttggaat tggccgcttc
     2701 actacagagg aggaagtgga ctacacagtg gagaaatgca ttcagcatgt gaagcgtctt
     2761 cgagaaatga gccctctctg ggagatggtt caggatggca ttgacctcaa gagcatcaag
     2821 tggacccaac actagaagaa tagggccctg agaattcgcc agcacagtgg tcgaccctgt
     2881 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2941 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3061 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3121 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3181 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tgccagctgg ggcgccctct
     3241 ggtaaggttg ggaagccctg cagatctgat cccggggggc aatgagatat gaaaaagcct
     3301 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac
     3361 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
     3421 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
     3481 cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaatttagc
     3541 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct
     3601 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg
     3661 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac
     3721 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
     3781 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
     3841 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     3901 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     3961 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     4021 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg
     4081 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     4141 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     4201 gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta
     4261 ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga atagagtaga
     4321 tgccgaccgg gatctatcga tgcggcgggt gtggtggtta cgcgtaatct gctgcttgca
     4381 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4441 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4501 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4561 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4621 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4681 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4741 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4801 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4861 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4921 cctatggaaa aacgccagca acgcatcgat aaaataaaag attttattta gtctccagaa
     4981 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     5041 ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac
     5101 agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     5161 ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     5221 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca
     5281 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
     5341 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
     5401 ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg
     5461 agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
     5521 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
     5581 acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac attaaatggc
     5641 catagtgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5701 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5761 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggcccacagt
     5821 gctgcaatga taccgcgaga ctttttgttt gcgagtcagc agattacgcg cagaaaaaaa
     5881 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5941 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttn
     6001 nnnnnnnnnn gmrgyygyaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6061 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6121 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6181 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6241 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6301 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6361 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6421 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6481 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6541 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6601 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6661 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6721 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6781 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6841 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6901 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagg
//
